Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Broth Medium D (Lactose Monohydrate Broth)
For detection of coliform bacteria in water, food, dairy products in accordance with EP/BP.
Gms/Lit : 12.75
39.22 Lit/500G
Gms/Lit : 12.75
39.22 Lit/500G
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Dehydrated Culture Media#4fa25a
Intended Use
Recommended for detection of coliform bacteria in water, food, dairy products in accordance with EP/BP.
Composition
| Ingredients | g/L |
|---|---|
| Gelatin peptone $ | 5.000 |
| HM peptone B # | 3.000 |
| Lactose monohydrate | 5.000 |
| pH after sterilization (at 25°C) | 6.9±0.2 |
**Formula adjusted, standardized to suit performance parameters
# Equivalent to Beef extract, $ Equivalent to Pancreatic digest of gelatin
Directions
Suspend 12.75 grams (the equivalent weight of dehydrated medium per litre) in 1000 ml purified/distilled water. Heat if necessary to dissolve the medium completely. For larger inocula (10 ml or more) concentrated medium may be prepared to account for medium dilution by the inoculum. Mix well and distribute into tubes with inverted Durham's tubes. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes and cool immediately.
Principle And Interpretation
Fluid Lactose Medium is recommended by EP, BP, APHA (1-3) in the performance and confirmation of the presumptive test for coliform bacteria in pharmaceutical products (1,2), water (3), food (4) and milk (5). This medium can be used as an alternate to Lauryl Sulphate Broth in the presumptive test of the MPN of standard coliforms. This medium is also used for pre-enrichment of Salmonella for its detection in pharmaceutical raw materials. Gelatin peptone and HM peptone B supply essential nutrients to the organisms. Lactose is a fermentable carbohydrate for the coliforms. Tubes of Fluid Lactose Medium are inoculated with dilutions of water or milk, etc. under test, and incubated at 35-37°C and examined for gas formation after 24 and 48 hours. Members of the coliform group are defined as aerobic and facultative anaerobic gram-negative and non-sporing bacilli which ferment lactose with gas formation within 48 hours at 35°C. In testing dairy products, Fluid Lactose Medium is used only in the completed test (4). Large water samples may require double strength Fluid Lactose Medium to minimize the final volume.
Type of specimen
Pharmaceutical samples; Water samples; Food and dairy samples
Specimen Collection and Handling
For pharmaceutical samples, follow appropriate techniques for sample collection, processing as per guidelines (1,2). For water samples, follow appropriate techniques for sample collection, processing as per guidelines and local standards (3). For food and dairy samples, follow appropriate techniques for sample collection and processing as per guidelines (4,5). After use, contaminated materials must be sterilized by autoclaving before discarding.
Warning and Precautions
Read the label before opening the container. Wear protective gloves/protective clothing/eye protection/ face protection. Follow good microbiological lab practices while handling specimens and culture. Standard precautions as per established guidelines must be followed while handling specimens. Safety guidelines may be referred in individual safety data sheets.
Limitations
- 1.Large water samples may require double strength Lactose Monohydrate Medium to minimize the final volume.
- 2.Further biochemical identification is necessary for confirmation.
Performance and Evaluation
Performance of the medium is expected when used as per the direction on the label within the expiry period when stored at recommended temperature.
Quality Control
Appearance
Cream to yellow homogeneous free flowing powder
Colour and clarity of prepared medium
Light amber coloured clear solution in tubes
pH
6.70-7.10
Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for 18-48 hours.
| Organism | Inoculum (CFU) |
Growth | Gas |
|---|---|---|---|
| #Klebsiella aerogenes ATCC 13048 (00175*) |
50-100 | luxuriant | Positive reaction |
| Enterococcus faecalis ATCC 29212 (00087*) |
50-100 | luxuriant | Negative reaction |
| ^Pseudomonas paraeruginosa ATCC 9027 (00026*) |
50-100 | luxuriant | Negative reaction |
| Escherichia coli ATCC 8739 (00012*) |
50-100 | luxuriant | Positive reaction |
| Salmonella Typhimurium ATCC 14028 (00031*) |
50-100 | luxuriant | Negative reaction |
| Salmonella Abony NCTC 6017 (00029*) |
50-100 | luxuriant | Negative reaction |
Key: * Corresponding WDCM numbers,
(#) Formerly known as Enterobacter aerogenes
(^) Formerly known as Pseudomonas aeruginosa
Storage and Shelf Life
Store between 10-30°C in tightly closed container and the prepared medium at 15-30°C. Use before expiry date on the label. On opening, product should be properly stored dry, after tightly capping the bottle in-order to prevent lump formation due to the hygroscopic nature of the product. Improper storage of the product may lead to lump formation. Store in dry ventilated area protected from extremes of temperature and sources of ignition. Seal the container tightly after use. Product performance is best if used within stated expiry period.
Disposal
User must ensure safe disposal by autoclaving and/or incineration of used or unusable preparations of this product. Follow established laboratory procedures in disposing of infectious materials and material that comes into contact with sample must be decontaminated and disposed of in accordance with current laboratory techniques (6,7).
References
- 1. European Pharmacopoeia, 2022, 10 th volume, European Directorate for the quality of medicines & Healthcare.
- 2. The British Pharmacopoeia, 2022, Medicines and Healthcare products Regulatory Agency.
- 3. Lipps WC, Braun-Howland EB, Baxter TE,eds. Standard methods for the Examination of Water and Wastewater, 24th ed. Washington DC:APHA Press; 2023.
- 4. Salfinger Y., and Tortorello M.L., 2015, Compendium of Methods for the Microbiological Examination of Foods, 5th Ed., American Public Health Association, Washington, D.C.
- 5. Wehr H M and Frank J H., 2004, Standard Methods for the Examination of Dairy Products, 17th ed., APHA Inc., Washington, D.C.
- 6. Isenberg, H.D. Clinical Microbiology Procedures Handbook. 2nd Edition.
- 7. Jorgensen, J.H., Pfaller, M.A., Carroll, K.C., Funke, G., Landry, M.L., Richter, S.S and Warnock., D.W. (2015) Manual of Clinical Microbiology, 11th Edition. Vol. 1.
| Product Name | Broth Medium D (Lactose Monohydrate Broth) |
|---|---|
| SKU | MAP1003 |
| Customized Product Available | No |
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